脐血浆培养的脐血树突状细胞对胃癌细胞的特异杀伤作用

The specific cytotoxic effects to gastric cancer cells induced by human cord blood dendritic cells cultured with human umbilical cord plasma

目的尝试用脐血浆代替胎牛血清培养脐血树突状细胞(DCs),并使之负载胃癌抗原,观察其对胃癌细胞的特异杀伤作用。方法分离脐血单个核细胞(CBMCs)并在含10%同源脐血浆的培养体系中诱导培养,部分细胞加入胃癌冻融抗原冲击。流式细胞仪检测细胞表面抗原CD1a和CD83表达,MTT法检测DCs体外刺激淋巴细胞增殖活性,LDH法检测DCs诱导的细胞毒T淋巴细胞(CTLs)对胃癌及肝癌细胞的细胞毒作用。结果CBMCs能分化为表型正常的未成熟DCs,并进而吞噬胃癌细胞冻融抗原成熟,刺激淋巴细胞增殖并诱导对胃癌细胞株特异的CTLs毒性。结论脐血浆培养的脐血DCs能有效捕获胃癌冻融抗原,激发针对胃癌细胞的特异杀伤效应。本实验采用的DCs制备方法具有方法简单、成本较低、瘤苗制备时间较短等优点。

Objective To evaluate the effect of replacing fetal cattle serum with umbilical cord plasma on incubating dendritic cells （DCs） from umbilical cord blood in vitro and observe the anti-tumor response to gastric cancer cells induced by DCs after impacted with freeze thawing antigen. Methods Human cord blood mononuclear cells （CBMCs） were cultured in RPMI1640 containing 10% autologous plasma, some of which were supplemented with tumor lyaates. The cells were shown by flow cytometry to express CD1a and CD83. MTT assay was applied to measure allostimulate capacity of DCs. Killing rates of autologous T lymphocytes induced by DCs on different target cells were measured by LDH method, Results Cells appeared typical phenotype of DCs after culture. The tumor lysate antigens could be captured by DCs and then presented to lymphocytes successfully to stimulate the proliferation. The cytotoxic T lymphocytes （CTLs） response in vitro was enhanced by treating with tumor lysates. Conclusion The DCs can capture the antigen utility and induce the specific cytotoxic effects to gastric cancer cells. The method to incubate DCs in this study is simple, low-cost and short-time.